RESUMO
One of the most cited limitations of capillary and microchip electrophoresis is the poor sensitivity. This review continues to update this series of biannual reviews, first published in Electrophoresis in 2007, on developments in the field of online/in-line concentration methods in capillaries and microchips, covering the period July 2016-June 2018. It includes developments in the field of stacking, covering all methods from field-amplified sample stacking and large-volume sample stacking, through to isotachophoresis, dynamic pH junction, and sweeping. Attention is also given to online or in-line extraction methods that have been used for electrophoresis.
Assuntos
Eletroforese Capilar , Animais , Biomarcadores/análise , Linhagem Celular , Fracionamento Químico , Humanos , Concentração de Íons de Hidrogênio , Isotacoforese , Camundongos , Micelas , Sensibilidade e EspecificidadeRESUMO
One of the most cited limitations of capillary (and microchip) electrophoresis is the poor sensitivity. This review continues to update this series of biennial reviews, first published in Electrophoresis in 2007, on developments in the field of on-line/in-line concentration methods in capillaries and microchips, covering the period July 2014-June 2016. It includes developments in the field of stacking, covering all methods from field amplified sample stacking and large volume sample stacking, through to isotachophoresis, dynamic pH junction, and sweeping. Attention is also given to on-line or in-line extraction methods that have been used for electrophoresis.
Assuntos
Eletroforese Capilar/métodos , Isotacoforese/métodos , Procedimentos Analíticos em Microchip/métodos , Eletrocromatografia Capilar , Linhagem Celular , Humanos , Compostos Inorgânicos/análise , Focalização Isoelétrica , Nanoestruturas/análise , Compostos Orgânicos/análise , Sensibilidade e EspecificidadeRESUMO
Counter-pressure was used to extend the duration of field-amplified sample injection in isotachophoresis (FASI-ITP) in order to improve the detection of bacterial cells. Using 0.51-µm negatively charged encapsulated fluorescent beads as a model, the counter-pressure, injection and separation voltages, and times were optimized. Using 6-min 8,963-Pa counter-pressure FASI-ITP injections at -12 kV followed by mobilization of the ITP band with continued injection at -6 kV, the limit of detection (LOD) for Escherichia coli was improved to 78 cells/mL, a factor of 4 when compared with FASI-ITP without counter-pressure.
Assuntos
Carga Bacteriana/métodos , Separação Celular/métodos , Eletroforese Capilar/métodos , Monitoramento Ambiental/métodos , Escherichia coli/isolamento & purificação , Isotacoforese/métodos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
One of the most cited limitations of capillary (and microchip) electrophoresis is the poor sensitivity. This review continues to update this series of biannual reviews, first published in Electrophoresis in 2007, on developments in the field of on-line/in-line concentration methods, covering the period July 2012-July 2014. It includes developments in the field of stacking, covering all methods from field-amplified sample stacking and large-volume sample stacking, through to ITP, dynamic pH junction, and sweeping. Attention is also given to on-line or in-line extraction methods that have been used for electrophoresis.
Assuntos
Eletroforese Capilar/instrumentação , Eletroforese Capilar/métodos , Animais , Desenho de Equipamento , Humanos , Concentração de Íons de Hidrogênio , Extração Líquido-Líquido/instrumentação , Extração Líquido-Líquido/métodos , Tamanho da Amostra , Sensibilidade e Especificidade , Extração em Fase Sólida/instrumentação , Extração em Fase Sólida/métodosRESUMO
A highly sensitive capillary isotachophoresis method with LIF detection for microbial analysis was developed. This allowed the reliable analysis of Escherichia coli bacteria with a LOD of 14 cells in a sample volume of 100 µL, or 1.35 × 10(2) cell/mL, which is 47 times lower than reported by CE-LIF and 148 times lower than CE-UV with on-line concentration. A leading electrolyte of 50 mM Tris-HCl was used while the cells were diluted in 5 mM Tris HEPES as the terminator. To facilitate detection, cells were stained with the universal nucleic acid fluorophore SYTO 9. Continuous electrokinetic injection of the cells from the terminator under field amplified conditions concentrated cells into a single peak at the leader/terminator boundary allowing quantitation by measurement of peak height. The method was applied to water collected from two local streams, with only filtration through a 5-µm syringe filter to remove large particulate matter followed by a ten times dilution in terminator, with total analysis time approximately 40 min. The detected cell numbers in the water samples by the isotachophoresis method were 3.70 × 10(5) cell/mL and 2.62 × 10(4) cell/mL, which were slightly higher than the 9.50 × 10(4) cell/mL and 1.96 × 10(4) cell/mL obtained by conventional microbiological plate counting.